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During 2,3,5,6-tetramethylpyrazine production from glucose by Bacillus strains, a novel product was detected and identified as 2,4,5-trimethylimidazole (TMI) by GC/MS. TMI appeared in the culture
medium only after glucose had been depleted and then increased to 0.25–0.31 g l−1 in 90–120 h. When the ammonium source was changed from (NH4)2SO4 to (NH4)2HPO4, only about one tenth of TMI was detected. Although the mechanistic events largely remain unclear, both microbial strains
tested demonstrated similar dynamic processes of TMI production, suggesting that TMI formation is a genuine feature of Bacillus species.
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